Difference between revisions of "Part:BBa K3792006"
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This generator was assembled into the pSB1K3 plasmid, and then transformed into E.Coli NEB 5-Alpha. To test if the cells were secreting chitin, The FSU 2021 team used the Chitosan Assay Kit (XAN-5126) available from Cell Biolabs. The manual for this kit contains the protocols for converting chitin to chitosan, along with a protocol for detecting chitosan. | This generator was assembled into the pSB1K3 plasmid, and then transformed into E.Coli NEB 5-Alpha. To test if the cells were secreting chitin, The FSU 2021 team used the Chitosan Assay Kit (XAN-5126) available from Cell Biolabs. The manual for this kit contains the protocols for converting chitin to chitosan, along with a protocol for detecting chitosan. | ||
− | Chitin from the cells was converted into chitosan by reacting with 12.5M NaOH at 95°C for 24 hours. The solution was then neutralized with acetic acid. The assay was then performed on the supernatant of the cell culture. | + | Chitin from the cells was converted into chitosan by reacting with 12.5M NaOH at 95°C for 24 hours. The solution was then neutralized with acetic acid. The assay was then performed on the supernatant of the cell culture. |
+ | |||
+ | <table id="modular-promoter-library-chart" class="charts-css column hide-data show-primary-axis show-10-secondary-axes"> | ||
+ | <tr> | ||
+ | <td style="--size: calc(897/900)"><span class="tooltip">iGEM: [https://parts.igem.org/Part:BBa_M36303 BBa_M36303]<br>BIOFAB: apFAB46<br>Strength: 897</span><span class="data">897</span></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="--size: calc(866.7/900)"><span class="tooltip">iGEM: [https://parts.igem.org/Part:BBa_K2832101 BBa_K2832101]<br>BIOFAB: apFAB70<br>Strength: 866.7</span><span class="data">866.7</span></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="--size: calc(866/900)"><span class="tooltip">iGEM: [https://parts.igem.org/Part:BBa_K2832102 BBa_K2832102]<br>BIOFAB: apFAB71<br>Strength: 866</span><span class="data">866</span></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="--size: calc(857/900)"><span class="tooltip">iGEM: [https://parts.igem.org/Part:BBa_K2832103 BBa_K2832103]<br>BIOFAB: apFAB61<br>Strength: 857</span><span class="data">857</span></td> | ||
+ | </tr> | ||
+ | </table> | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Revision as of 00:44, 22 October 2021
NodCIJ Generator I
The NodCIJ Generator with a weak promoter does not produce a lot of the NodC protein, NodI protein, and NodJ protein. The NodC protein produces chitin and NodI and NodJ transports nodulation factors.
This part consists of 2555 base-pair length. It includes a promoter, 3 ribosome binding sites, 3 protein-coding sequences, and a terminator. We used promoter K2832184, a weak promoter from the BIOFAB collection. In addition, we also used protein-coding sequences from Chitin synthase from Rhiozobium leguminosarum, Nodl a protein possibly involved in transporting chitin across the cell membrane, and NodJ, a protein also possibly involved in the transportation of chitin
This generator was assembled into the pSB1K3 plasmid, and then transformed into E.Coli NEB 5-Alpha. To test if the cells were secreting chitin, The FSU 2021 team used the Chitosan Assay Kit (XAN-5126) available from Cell Biolabs. The manual for this kit contains the protocols for converting chitin to chitosan, along with a protocol for detecting chitosan.
Chitin from the cells was converted into chitosan by reacting with 12.5M NaOH at 95°C for 24 hours. The solution was then neutralized with acetic acid. The assay was then performed on the supernatant of the cell culture.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 492
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 854